A temperature-controlled plant psychrometer

When measuring plant water potential in conditions of fluctuating temperature, commercial psychrometers exhibit large errors which are caused by a temperature difference between the reference junctions and the chamber air. A novel psychrometer capable of field use is described which incorporates an electronic device for controlling reference junction temperatures. It consists of a chamber with a thermocouple mounted in it for measuring dewpoints and another thermocouple for sensing the temperature gradients. A thermo-electric heat-pump keeps the temperature of the reference junctions the same as that of the chamber air. Good agreement was found between measurements made with this psychrometer and independent measurements made with the destructive pressure chamber method in the glasshouse and in the field under conditions of widely fluctuating temperature. Corresponding measurements with a commercial psychrometer without temperature control failed to show such agreement. Heavy thermal insulation is necessary, despite the temperature control, which can restrict the psychrometers used. Possibilities for further improvements are discussed.     

Plant water potential measured continuously in the field

Summary A simple system is described which allowed the use of a stem attached psychrometer in the field. Water was circulated between a tank buried in the soil and the psychrometer and this sytem reduced temperature gradients sufficiently to enable continuous measurements of plant water potential to be made in the field. Measurements agreed with those taken independently with a destructive pressure chamber method.     

Leaf water potentials measured with a pressure chamber

Leaf water potentials were estimated from the sum of the balancing pressure measured with a pressure chamber and the osmotic potential of the xylem sap in leafy shoots or leaves. When leaf water potentials in yew, rhododendron, and sunflower were compared with those measured with a thermocouple psychrometer known to indicate accurate values of leaf water potential, determinations were within ± 2 bars of the psychrometer measurements with sunflower and yew. In rhododendron. water potentials measured with the pressure chamber plus xylem sap were 2.5 bars less negative to 4 bars more negative than psychrometer measurements.

The discrepancies in the rhododendron measurements could be attributed, at least in part, to the filling of tissues other than xylem with xylem sap during measurements with the pressure chamber. It was concluded that, although stem characteristics may affect the measurements, pressure chamber determinations were sufficiently close to psychrometer measurements that the pressure chamber may be used for relative measurements of leaf water potentials, especially in sunflower and yew. For accurate determinations of leaf water potential, however, pressure chamber measurements must be calibrated with a thermocouple psychrometer.

     

In situ field measurement of leaf water potential using thermocouple psychrometers

Thermocouple psychrometers are the only instruments which can measure the in situ water potential of intact leaves, and which can possibly be used to monitor leaf water potential. Unfortunately, their usefulness is limited by a number of difficulties, among them fluctuating temperatures and temperature gradients within the psychrometer, sealing of the psychrometer chamber to the leaf, shading of the leaf by the psychrometer, and resistance to water vapor diffusion by the cuticle when the stomates are closed. Using Citrus jambhiri, we have tested several psychrometer design and operational modifications and showed that in situ psychrometric measurements compared favorably with simultaneous Scholander pressure chamber measurements on neighboring leaves when the latter were corrected for the osmotic potential.     

Comparison of the dye method with the thermocouple psychrometer for measuring leaf water potentials

The dye method for measuring water potential was examined and compared with the thermocouple psychrometer method in order to evaluate its usefulness for measuring leaf water potentials of forest trees and common laboratory plants. Psychrometer measurements are assumed to represent the true leaf water potentials. Because of the contamination of test solutions by cell sap and leaf surface residues, dye method values of most species varied about 1 to 5 bars from psychrometer values over the leaf water potential range of 0 to −30 bars. The dye method is useful for measuring changes and relative values in leaf potential. Because of species differences in the relationships of dye method values to true leaf water potentials, dye method values should be interpreted with caution when comparing different species or the same species growing in widely different environments. Despite its limitations the dye method has a usefulness to many workers because it is simple, requires no elaborate equipment, and can be used in both the laboratory and field.     

A low-cost psychrometer for field measurements of atmospheric humidity

A design for an aspirated wet and dry bulb psychrometer, suitable for continuous field measurement of atmospheric humidity, is described. The psychrometer is simple and cheap to make, with a material cost of UK £40 per unit. It is mainly constructed from ready-made plastic plumbing parts, thus avoiding the need for expensive workshop facilities. The wet and dry bulb temperature sensors are suitable for direct connection to a solid-state data-logger. During extensive field use, it has proved reliable and easy to maintain. The accuracy of the device was assessed by intercomparison with an Assman psychrometer, a standard instrument for measuring atmospheric humidity. The test was conducted under high ambient saturation deficits (4.08 ± 1.22 kPa) and shortwave radiation fluxes in Niger, West Africa. Reasonable agreement between the two instruments was observed, with the low-cost psychrometer giving a mean saturation deficit 0.12 ± 0.25 kPa higher than the Assman.     

Pressure probe and isopiestic psychrometer measure similar turgor

Turgor measured with a miniature pressure probe was compared to that measured with an isopiestic thermocouple psychrometer in mature regions of soybean (Glycine max [L.] Merr.) stems. The probe measured turgor directly in cells of intact stems whereas the psychrometer measured the water potential and osmotic potential of excised stem segments and turgor was calculated by difference. When care was taken to prevent dehydration when working with the pressure probe, and diffusive resistance and dilution errors with the psychrometer, both methods gave similar values of turgor whether the plants were dehydrating or rehydrating. This finding, together with the previously demonstrated similarity in turgor measured with the isopiestic psychrometer and a pressure chamber, indicates that the pressure probe provides accurate measurements of turgor despite the need to penetrate the cell. On the other hand, it suggests that as long as precautions are taken to obtain accurate values for the water potential and osmotic potential, turgor can be determined by isopiestic psychrometry in tissues not accessible to the pressure probe for physical reasons.     

Effect of Cuticular Abrasion on Thermocouple Psychrometric In Situ Measurement of Leaf Water Potential

Cuticular resistance to water vapour diffusion between the substomatalcavity and the sensing psychrometer junction is a problem uniqueto leaf hygrometry. This resistance is not encountered in soilor solution hygrometry. The cuticular resistance may introduceerror in the measurement of leaf water potential. Using in situleaf hygrometers, we studied the effect of abrading the cuticleof Citrus jambhiri Lushington leaves, to reduce the diffusiveresistance. Field measurements of psychrometer water potentialwere compared with Scholander pressure chamber values for adjacentleaves. Different treatments were compared by sealing pairsof psychrometers on either side of the midrib. The time forwater vapour equilibration between the leaf and the psychrometerchamber was greater than 5 h for no abrasion. For abraded leaves,the true water potential value was obtained within an hour.After equilibration, psychrometer values compared favourablywith pressure chamber values for adjacent leaves (r > 0.97).Measured water potential for unabraded leaves did not correlatewell with corresponding pressure chamber measurements. Scanning electron micrographs indicated that the damage causedby abrading leaves for 60 s using carborundum powder (60 µmdiameter) was surface localized, with numerous scratchings ofthe leaf cuticle. The coarse abrasion treatment (aluminium oxide,75 µm diameter) resulted in fewer but larger cavitiesin the epidermis, which may explain the observed variabilityin the corresponding psychrometric measurements. Key words: Leaf water potential, Cuticular resistance, Leaf abrasion, Thermocouple psychrometer     

Psychrometric Field Measurement of Water Potential Changes following Leaf Excision

In situ measurement of sudden leaf water potential changes has not been performed under field conditions. A laboratory investigation involving the measurement of leaf water potential prior to and 2 to 200 minutes after excision of citrus leaves (Citrus jambhiri) showed good linear correlation (r = 0.99) between in situ leaf psychrometer and Scholander pressure chamber measurements. Following this, a field investigation was conducted which involved psychrometric measurement prior to petiole excision and 1 minute after excision. Simultaneous pressure chamber measurements were performed on neighboring leaves prior to the time of excision and then on the psychrometer leaf about 2 minutes after excision. These data indicate that within the first 2 minutes after excision, psychrometer and pressure chamber measurements were linearly correlated (r = 0.97). Under high evaporative demand conditions, the rate of water potential decrease was between 250 and 700 kilopascals in the first minute after excision. These results show that the thermocouple psychrometer can be used as a dynamic and nondestructive field technique for monitoring leaf water potential.     

Theoretical and experimental errors for in situ measurements of plant water potential

Errors in psychrometrically determined values of leaf water potential caused by tissue resistance to water vapor exchange and by lack of thermal equilibrium were evaluated using commercial in situ psychrometers (Wescor Inc., Logan, UT) on leaves of Tradescantia virginiana (L.). Theoretical errors in the dewpoint method of operation for these sensors were demonstrated. After correction for these errors, in situ measurements of leaf water potential indicated substantial errors caused by tissue resistance to water vapor exchange (4 to 6% reduction in apparent water potential per second of cooling time used) resulting from humidity depletions in the psychrometer chamber during the Peltier condensation process. These errors were avoided by use of a modified procedure for dewpoint measurement. Large changes in apparent water potential were caused by leaf and psychrometer exposure to moderate levels of irradiance. These changes were correlated with relatively small shifts in psychrometer zero offsets (−0.6 to −1.0 megapascals per microvolt), indicating substantial errors caused by nonisothermal conditions between the leaf and the psychrometer. Explicit correction for these errors is not possible with the current psychrometer design.     

An evaluation of techniques for measuring yield turgor in excised Salix leaves

Abstract. The use of three techniques for determining yield turgor in excised Salix leaves was investigated. These were the osmotic-solutions technique, the psychrometer technique, and the pressure-chamber technique. The application of the osmotic-solutions technique to a range of leaf types was discussed and the appropriate corrections for volume changes and the contribution of apoplastic water were detailed. It was concluded that the osmotic-solutions technique is not satisfactory for use with slowly growing and/or very elastic leaves. The psychrometer and pressure-chamber techniques were both simple compared with the osmotic-solutions technique, and gave values for yield turgor in the range of 0·3–0·5 MPa. A disadvantage of the psychrometer technique for field applications is that it requires one psychrometer chamber per sample. The pressure-chamber technique was modified for use as a field technique where multiple sampling could be easily and inexpensively achieved. Particular care was required with this technique to prevent water loss from the leaf during stress relaxation, but simple and effective procedures for doing so were found. The modified pressure-chamber technique described here, is recommended as the preferred technique for measuring the yield turgor of leaves in experiments where many simultaneous estimates of yield turgor are to be made.     

Errors in measuring water potentials of small samples resulting from water adsorption by thermocouple psychrometer chambers

The adsorption of water by thermocouple psychrometer assemblies is known to cause errors in the determination of water potential. Experiments were conducted to evaluate the effect of sample size and psychrometer chamber volume on measured water potentials of leaf discs, leaf segments, and sodium chloride solutions. Reasonable agreement was found between soybean (Glycine max L. Merr.) leaf water potentials measured on 5-millimeter radius leaf discs and large leaf segments. Results indicated that while errors due to adsorption may be significant when using small volumes of tissue, if sufficient tissue is used the errors are negligible. Because of the relationship between water potential and volume in plant tissue, the errors due to adsorption were larger with turgid tissue. Large psychrometers which were sealed into the sample chamber with latex tubing appeared to adsorb more water than those sealed with flexible plastic tubing. Estimates are provided of the amounts of water adsorbed by two different psychrometer assemblies and the amount of tissue sufficient for accurate measurements of leaf water potential with these assemblies. It is also demonstrated that water adsorption problems may have generated low water potential values which in prior studies have been attributed to large cut surface area to volume ratios.     

Improved Thermocouple Psychrometer for the Measurement of Plant and Soil Water Potential: III. EQUILIBRATION

The equilibration between the water potential of air insidethe psychrometer and that of calibrating solutions, soils, andplant leaves is examined. The results are compared with thosein the literature and with a theory for predicting equilibrationtimes. Techniques aimed at reducing equilibration times aredevised and tested. A method which employs a psychrometer todetermine the adsorption isotherm of its own wall material isused to assess the suitability of three materials for chamberconstruction. The main findings include: (1) Times for equilibration of solutionand soil samples were as brief as 8 min and 1–3 h, respectively,and times for ‘equilibration’ of tobacco leaf samplesobtained with the improved psychrometer are significantly shorterthan those reported for previous designs of Spanner-type psychrometers.These improvements enhance the calibration and operational convenienceof the method. (2) Current criteria for deciding the attainmentof "equilibria" with leaf samples may result in a large error,termed equilibration error. With pine and wheat leaf samplesthis error may be more than –460 J kg^–1, and about–1200 J kg^–1, respectively. A criterion for reducingequilibration error is described. (3) Predicted equilibrationtimes for leaf samples are generally far too short. Severalfactors that are neglected in the theory and which probablyaccount for most of this discrepancy are described. (4) A newtechnique is outlined which enables up to a 15-fold reductionin the ‘equilibration’ times of pine needle samples.(5) Stainless steel (type 316) appears to be the most suitablechamber material of those tested.     

Phloem water relations and translocation

Satisfactory measurements of phloem water potential of trees can be obtained with the Richards and Ogata psychrometer and the vapor equilibration techniques, although corrections for loss of dry weight and for heating by respiration are required for the vapor equilibrium values. The psychrometer technique is the more satisfactory of the 2 because it requires less time for equilibration, less tissue, and less handling of tissue. Phloem water potential of a yellow-poplar tree followed a diurnal pattern quite similar to that of leaves, except that the values were higher (less negative) and changed less than in the leaves.

The psychrometer technique permits a different approach to the study of translocation in trees. Measurements of water potential of phloem discs followed by freezing of samples and determination of osmotic potential allows estimation of turgor pressure in various parts of trees as the difference between osmotic potential and total water potential. This technique was used in evaluating gradients in water potential, osmotic potential, and turgor pressure in red maple trees. The expected gradients in osmotic potential were observed in the phloem, osmotic potential of the cell sap increasing (sap becoming more dilute) down the trunk. However, values of water potential were such that a gradient in turgor pressure apparently did not exist at a time when rate of translocation was expected to be high. These results do not support the mass flow theory of translocation favored by many workers.

     

The Relationship Between Leaf Thickness and Plant Water Potential

Leaf thickness was continuously measured in a wide range ofenvironments using a new type of displacement transducer whichis easy to set-up and automatically compensates for the effectsof temperature. Simultaneous measurements were made of waterpotential using either a psychrometer attached to the leaf petioleor a leaf pressure chamber. Thickness of leaves was a sensitiveindicator of plant water status but calibrations against anindependent method were necessary in every plant for accurateestimates of water potential. The relationship between leafthickness changes and water potential, measured in detachedleaves, was usually curvilinear and was strongly influencedby leaf age, stress history and, in young leaves, by the effectsof leaf growth. Leaf thickness growth was absent in mature cabbageleaves. Key words: Leaf thickness, plant water potential, psychrometer     

Chloroplast response to low leaf water potentials: I. Role of turgor

The effect of decreases in turgor on chloroplast activity was studied by measuring the photochemical activity of intact sunflower (Helianthus annuus L. cv. Russian Mammoth) leaves having low water potentials. Leaf turgor, calculated from leaf water potential and osmotic potential, was found to be affected by the dilution of cell contents by water in the cell walls, when osmotic potentials were measured with a thermocouple psychrometer. After the correction of measurements of leaf osmotic potential, both the thermocouple psychrometer and a pressure chamber indicated that turgor became zero in sunflower leaves at leaf water potentials of −10 bars. Since most of the loss in photochemical activity occurred at water potentials below −10 bars, it was concluded that turgor had little effect on the photochemical activity of the leaves.     

Improved Thermocouple Psychrometer for the Measurement of Plant and Soil Water Potential: II. OPERATION AND CALIBRATION

The operation characteristics (the Peltier cooling current iand the cooling period t_c), the range of measurement of waterpotential, the calibration sensitivity, and the accuracy areevaluated for an improved psychrometer and compared with datain the literature, and with a recent theory. The range increases with increasing Peltier cooling currentup to the optimal value of i 9 mA and has been found to agreefairly well with the predictions of theory. A maximum rangeof 0 to about –13 000 J/kg is obtainable with the improvedpsychrometer. This is nearly twice that previously reportedfor a Spanner-type psychrometer. The experimental calibration sensitivities C'_s increase withincreasing water potential, cooling period, and temperatureand, with t_c = 60 s, are in general larger than the theoreticalvalues at high water potentials but less than the theoreticalvalues at low water potentials. They are mostly larger thanthose reported by Monteith and Owen but about the same as thoseof Lang and Trickett. The accuracy of the improved psychrometer is significantly greaterthan that previously reported for a Spanner-type psychrometer. While the theory predicts a linear relationship between waterpotential and peak output, the observed calibration curves arenon-linear and vary between psychrometers. Because of thesediscrepancies between observation and theory, calibration ofeach psychrometer is necessary.     

Direct Demonstration of a Growth-Induced Water Potential Gradient

When transpiration is negligible, water potentials in growing tissues are less than those in mature tissues and have been predicted to form gradients that move water into the enlarging cells. To determine directly whether the gradients exist, we measured water potentials along the radius of stems of intact soybean (Glycine max [L.] Merr.) seedlings growing in vermiculite in a water-saturated atmosphere. The measurements were made in individual cells by first determining the turgor with a miniature pressure probe, then determining the osmotic potential of solution from the same cell, and finally summing the two potentials. The osmotic potentials were corrected for sample mixing in the probe. The measurements were checked with a thermocouple psychrometer that gave average tissue water potentials. In the elongating region, the water potential was highest near the xylem and lowest near the epidermis and in the center of the pith. In the basal, more mature region of the same stems, water potentials were near zero next to the xylem and throughout the tissue. These basal potentials reflected mostly the potential of the xylem, which extended into the elongating tissues. Thus, the high basal potential confirmed the high potential near the xylem in the elongating tissues. The psychrometer measurements for each tissue gave average potentials that agreed with the average of the cell potentials from the pressure probe. We conclude that a radial gradient was present in the elongating region that formed a water potential field in three dimensions around the xylem and that confirmed the predictions of Molz and Boyer (F.J. Molz and J.S. Boyer [1978] Plant Physiol 62: 423-429).     

Water potential gradient in a tall sequoiadendron

With an elevator installed in a 90-meter tall Sequoiadendron to collect the samples, xylem pressure potential measurements were made approximately every 15 meters along 60 meters of the tree's height. The measured gradient was about −0.8 bar per 10 meters of height, i.e., less than the hydrostatic gradient. Correction of the xylem pressure potential data by calibration against a thermocouple psychrometer confirmed this result. Similar gradients are described in the literature in tall conifers at times of low transpiration, although a different sampling technique was used. If the data in the present study and those supporting it are typical, they imply a re-evaluation of either the use of the pressure chamber to estimate water potential or the present theories describing water transport in tall trees.     

Field measurement of leaf water potential with a temperature-corrected in situ thermocouple psychrometer

Abstract. The construction and evaluation of a temperature-corrected in situ thermocouple psychrometer for measurement of leaf water potential (Ψ) is described. The instrument utilized two chromel-constantan thermocouples which allowed for detection of both the psychrometric zero offset and the temperature differential between the sample and the Peltier measuring junction. The psychrometer was subjected to stable temperature gradients while in contact with reference solutions of sodium chloride, and the effects of thermal gradients were quantified. Regression analysis indicated that temperature differentials were responsible for errors in water potential determinations of approximately –7.73 MPa°C⁻¹. When installed on leaves of field-grown cotton ( Gossypium hirsutum L.), corn ( Zea mays L.) and soybean ( Glycine max L. Merr) the instrument detected temperature differentials up to 0.1°C (–6.0 μV) which were associated with relatively small shifts in psychrometric zero offsets (–0.05––0.75 μV). Results indicated that substantial errors in apparent Ψ were caused by non-isothermal conditions between the leaf and the psychrometer measuring junction. The relative magnitude of these errors could be quantified and the corrected results showed good agreement with conventional psychrometric determination of Ψ on excised samples during a diurnal cycle.